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To explore a method for monitoring the concentration of SARS-CoV-2 in sewage in Shenzhen using digital PCR, enabling real-time detection of viral nucleic acids in sewage and reflecting the epidemiological status of COVID-19 infections in the population through nucleic acid concentration.
A digital PCR detection system for SARS-CoV-2 in sewage was established, and its minimum detection limit was determined. From April to July 2023, 24-hour composite sewage samples were collected twice weekly from the inflow of six sewage treatment plants in Nanshan and Futian districts of Shenzhen. The modified polyethylene glycol precipitation method was used for viral concentration enrichment, followed by quantitative detection of SARS-CoV-2 nucleic acids using the established reverse transcription digital PCR (RT-dPCR) method. Concurrently, data on COVID-19 infection cases in the monitored sewage areas were collected for correlation analysis with the nucleic acid concentrations in the sewage.
The RT-dPCR method for SARS-CoV-2 nucleic acid detection was successfully established, with a minimum detection limit of 1.00 copies/μl. A total of 162 sewage samples were collected from April to July 2023, with a positive detection rate of 96.3% for SARS-CoV-2 nucleic acids. The concentration of SARS-CoV-2 nucleic acids in Nanshan district ranged from 1.00×103 to 1.08×106 copies/L, while in Futian district it ranged from 1.44×103 to 1.40×106 copies/L, with peak concentrations observed on May 16. Correlation analysis indicated a significant association between sewage SARS-CoV-2 nucleic acid concentrations and the number of COVID-19 infection cases in the population (Nanshan district r=0.77, P<0.001; Futian district r=0.80, P<0.001).
The digital PCR method is suitable for the quantitative detection of low concentrations of SARS-CoV-2 nucleic acids in sewage, allowing for real-time monitoring of the COVID-19 epidemic, thus providing a scientific basis for public health management and intervention.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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